The non-catalytic function of PARP2 in DNA repair and cancer therapy
PARP2在DNA修复和癌症治疗中的非催化功能
基本信息
- 批准号:10540084
- 负责人:
- 金额:$ 59.71万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-07-01 至 2027-06-30
- 项目状态:未结题
- 来源:
- 关键词:ADP Ribose TransferasesADP ribosylationAcute Myelocytic LeukemiaAddressAdultAffectAffinityAnemiaAnimalsBRCA deficientBRCA1 geneBRCA2 geneBase Excision RepairsBindingBinding ProteinsBiochemicalBiochemistryBiological AssayBone MarrowCHEK2 geneCancer PatientCellsCellular biologyChargeChromatinChronicCisplatinClinicalClonal ExpansionDNADNA BindingDNA DamageDNA LigasesDNA RepairDNA Repair GeneDNA biosynthesisDNA lesionDefectDevelopmentDoseDysplasiaEmbryoErythropoiesisFlow CytometryFluorescenceGenesGenetic EpistasisGoalsHematopoiesisHematopoieticHistonesHypersensitivityImpairmentIn VitroKnock-inKnockout MiceLasersLigaseLigationLinkMalignant neoplasm of ovaryMalignant neoplasm of pancreasMalignant neoplasm of prostateMediatingModelingMolecular BiologyMusMutationMyelogenousOkazaki fragmentsPatientsPhotobleachingPhysical condensationPoly Adenosine Diphosphate RiboseProteinsRelaxationRiskRoleS phaseSiteSpecificitySplenomegalyTP53 geneTestingToxic effectWithdrawalXRCC1 genebasebrca genecancer cellcancer therapychemotherapyeffective therapyin vivoinhibitorlive cell imagingmalignant breast neoplasmmouse geneticsmouse modelnovelprematurepreventrecruitrepaired
项目摘要
PROJECT SUMMARY/ABSTRACT
PARP1 and PARP2 are DNA damage-induced poly-ADP-ribose (PAR) transferases, which are recruited
to and activated by DNA breaks. Active PARP1&2 PARylate themselves and histones to promote DNA repair
and chromatin relaxation. Dual-specificity inhibitors (PARPi) for PARP1 and PARP2 are currently used for the
treatment of BRCA1/2-deficient breast, ovarian, pancreatic, and prostate cancers. However, severe anemia
occurs in ~ one-third of patients, leading to dose reduction and premature termination of PAPRi therapy. PARPi
also cause significant clonal hematopoiesis, a condition that increases the risk for myeloid dysplasia and acute-
myeloid leukemia (MDS/AML). These hematopoietic toxicities are unexpected since the complete loss of Parp1
that is responsible for >80% of DNA damage-induced PARylation in cells, has no impact on hematopoiesis. To
understand this potential on-target toxicity of PARPi, we generated mouse models with knockin catalytically
inactive mutations in Parp2 – Poly-ADP-Ribosylation (PARylation) deficient (E534A, Parp2EA) or Mono-ADP-
Ribosylation (MARylation) deficient (H404A, Parp2HA). In contrast to the normal development of Parp2-/- mice,
mice expressing PARylation deficient PARP2 (Parp2EA/EA) died at embryonic day 16.5 (E16.5) with severe
anemia and stage-specific blocks in erythropoiesis. The Parp2HA/HA mice are viable but display splenomegaly
and chronic anemia. Meanwhile, our cell biology analyses suggest that clinically used PARPi effectively stalls
PARP2, but not PARP1, on laser-induced DNA damage sites. Based on these and other observations, we
hypothesize that PARP2 has PARylation-dependent structural functions that preferentially block nick
ligation during DNA replication, underlie the PARP inhibitors-induced erythropoiesis defects and anemia.
Specifically, we propose that catalytically inactive PARP2 were trapped on DNA breaks, especially 5’pho-nicks,
where they prevent other repair factors (e.g., Ligase1) from accessing the DNA nicks. Correspondingly, like the
Parp2EA/EA mice, Lig1-/- mice also succumbed to lethal anemia at E16.5. To test our hypothesis, we will Aim 1)
investigate the mechanism that regulates PARP2 recruitment and dynamics at DNA damage sites using live-cell
imaging and bulk biochemical assays; Aim 2) determine the impacts of catalytically inactive PARP2 on the
recruitment and function of other DNA repair factors, and DNA replication in vivo and in vitro; Aim 3) characterize
the mouse models expressing catalytically inactive - Parp2 and investigate how the loss of Trp53, CHK2, two
genes associated clonal hematopoiesis, modulates the anemia in Parp2EA/EA models. The completion of the
proposal will characterize the previously unrecognized structural functions of PARP2, address how catalytically
inactive PARP2 compromises DNA replication and selectively abrogates erythropoiesis, providing the strategy
to mitigate this on-targeted toxicity of PARP inhibitors by reducing PARP2 stalling.
项目总结/摘要
PARP 1和PARP 2是DNA损伤诱导的多聚ADP核糖(PAR)转移酶,其被募集
并被DNA断裂激活活性PARP 1和2 PARylate自身和组蛋白以促进DNA修复
和染色质松弛。PARP 1和PARP 2的双特异性抑制剂(PARPi)目前用于治疗糖尿病。
治疗BRCA 1/2缺陷的乳腺癌、卵巢癌、胰腺癌和前列腺癌。然而,严重贫血
在约三分之一的患者中发生,导致剂量减少和PAPRi治疗的过早终止。PARPi
也会导致显著的克隆性造血,这种情况会增加骨髓发育不良和急性-
骨髓性白血病(MDS/AML)。这些造血毒性是出乎意料的,因为Parp 1的完全丧失
其负责细胞中>80%的DNA损伤诱导的PAR化,对造血没有影响。到
为了理解PARPi的这种潜在的靶向毒性,我们用催化敲入产生了小鼠模型,
Parp 2-聚ADP-核糖基化(PAR化)缺陷(E534 A,Parp 2 EA)或单ADP-核糖基化(PAR化)缺陷(E534 A,Parp 2 EA)中的无活性突变
核糖基化(MAR化)缺陷(H404 A,Parp 2 HA)。与Parp 2-/-小鼠的正常发育相反,
表达PAR化缺陷型PARP 2(Parp 2 EA/EA)的小鼠在胚胎第16.5天(E16.5)死亡,
贫血和红细胞生成阶段特异性阻滞。Parp 2 HA/HA小鼠存活,但显示脾肿大
和慢性贫血。同时,我们的细胞生物学分析表明,临床使用的PARPi有效地阻止了
PARP 2,而不是PARP 1,在激光诱导的DNA损伤位点。根据这些和其他观察,我们
假设PARP 2具有优先阻断切口PAR化依赖性结构功能
DNA复制过程中的连接,是PARP介导的红细胞生成缺陷和贫血的基础。
具体地说,我们提出,无催化活性的PARP 2被困在DNA断裂,特别是5 'pho-nicks,
其中它们阻止其它修复因子(例如,连接酶1)进入DNA缺口。相应地,如
Parp 2 EA/EA小鼠、Lig 1-/-小鼠也在E16.5死于致死性贫血。为了验证我们的假设,我们将目标1)
研究使用活细胞在DNA损伤位点调节PARP 2募集和动力学的机制
成像和大量生物化学测定;目的2)确定无催化活性的PARP 2对细胞的影响,
其他DNA修复因子的募集和功能,以及体内和体外的DNA复制;目的3)表征
表达无催化活性的-Parp 2的小鼠模型,并研究Trp 53,CHK 2,两个
与克隆造血相关的基因调节Parp 2 EA/EA模型中的贫血。的完成
一项提案将描述PARP 2以前未被认识到的结构功能,
失活的PARP 2损害DNA复制并选择性地消除红细胞生成,
以通过减少PARP 2停滞来减轻PARP抑制剂的这种靶向毒性。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Shan Zha其他文献
Shan Zha的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Shan Zha', 18)}}的其他基金
The role of DNA-PKcs in DNA repair, lymphocyte development, RNA metabolism and tumor suppression
DNA-PKcs 在 DNA 修复、淋巴细胞发育、RNA 代谢和肿瘤抑制中的作用
- 批准号:
10539944 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
The role of DNA-PKcs in DNA repair, lymphocyte development, RNA metabolism and tumor suppression
DNA-PKcs 在 DNA 修复、淋巴细胞发育、RNA 代谢和肿瘤抑制中的作用
- 批准号:
10651884 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
The non-catalytic function of PARP2 in DNA repair and cancer therapy
PARP2在DNA修复和癌症治疗中的非催化功能
- 批准号:
10641934 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
The catalytic and non-catalytic functions of PARP1 in cancer biology
PARP1 在癌症生物学中的催化和非催化功能
- 批准号:
10377548 - 财政年份:2018
- 资助金额:
$ 59.71万 - 项目类别:
The catalytic and non-catalytic functions of PARP1 in cancer biology
PARP1 在癌症生物学中的催化和非催化功能
- 批准号:
9886208 - 财政年份:2018
- 资助金额:
$ 59.71万 - 项目类别:
The structural function of ATR in development, oncogenesis and cancer therapy
ATR 在发育、肿瘤发生和癌症治疗中的结构功能
- 批准号:
9886205 - 财政年份:2017
- 资助金额:
$ 59.71万 - 项目类别:
DNA-PKCS Phosphorylation in DNA Repair and Chromosomal Translocations
DNA 修复和染色体易位中的 DNA-PKCS 磷酸化
- 批准号:
8975763 - 财政年份:2014
- 资助金额:
$ 59.71万 - 项目类别:
相似海外基金
Control of genomic integrity and virulence of Aspergillus fumigatus by ADP-ribosylation.
通过 ADP-核糖基化控制烟曲霉的基因组完整性和毒力。
- 批准号:
MR/X007472/1 - 财政年份:2023
- 资助金额:
$ 59.71万 - 项目类别:
Fellowship
Understanding the impact of DNA ADP-ribosylation on telomere function in cancer cells
了解 DNA ADP-核糖基化对癌细胞端粒功能的影响
- 批准号:
10751121 - 财政年份:2023
- 资助金额:
$ 59.71万 - 项目类别:
Composition and function of telomeric multi-protein complexes and their regulation by ADP-ribosylation
端粒多蛋白复合物的组成和功能及其ADP-核糖基化的调节
- 批准号:
2748032 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
Studentship
A Chemical Footprinting Approach towards Poly-ADP-Ribosylation-regulated Biomolecular Condensation
聚 ADP 核糖基化调节生物分子缩合的化学足迹方法
- 批准号:
10524783 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
Regulation of DNA repair by histone ADP-ribosylation
组蛋白 ADP 核糖基化调节 DNA 修复
- 批准号:
MR/W017350/1 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
Research Grant
Regulation and function of site-specific protein poly-ADP-ribosylation
位点特异性蛋白质聚 ADP 核糖基化的调控和功能
- 批准号:
10668492 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
ADP-ribosylation of DNA in Mycobacterium tuberculosis
结核分枝杆菌 DNA 的 ADP-核糖基化
- 批准号:
BB/W016613/1 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
Research Grant
A Chemical Footprinting Approach towards Poly-ADP-Ribosylation-regulated Biomolecular Condensation
聚 ADP 核糖基化调节生物分子缩合的化学足迹方法
- 批准号:
10610165 - 财政年份:2022
- 资助金额:
$ 59.71万 - 项目类别:
A Chemical Footprinting Approach towards Poly-ADP-Ribosylation-regulated Biomolecular Condensation
聚 ADP 核糖基化调节生物分子缩合的化学足迹方法
- 批准号:
10389853 - 财政年份:2021
- 资助金额:
$ 59.71万 - 项目类别:
Role of Transcription Factor ADP-ribosylation in Breast Cancer Biology
转录因子 ADP-核糖基化在乳腺癌生物学中的作用
- 批准号:
10593900 - 财政年份:2021
- 资助金额:
$ 59.71万 - 项目类别: